This proposal seeks to develop a system for metabolic profiling of 3D Micro-Tissue (MT) in moderate throughput, and its testing on three complex MTs models of lung cancer (NSCLC) that will serve as predictive tools for response to inhibition of the Epidermal Growth Factor Receptor (EGFR) with erlotinib. Models will be formed from H3225 and PC9 NSCLC cells that harbor two of the most common EGFR activating mutations, L858R and exon 19 deletion respectively. As resistant controls, A549 and PC9GR NSCLC will be used. In Aim 1, a 96 well platform will be developed that facilitates spheroid formation from cell lines and disassociated NSCLC tumor tissue in one step, and these preps can be directly assayed for metabolism, toxicity and histology. Aim 2 will provide a mechanism by which intact MTs can be generated from incisional and core needle biopsies. Intact MTs will provide information on the metabolic and toxicity level, while maintaining in-vivo spatial information, cell-cell interactions and stromal influence. In Aim 3 the efficiency of predicting erlotonib toxicity will be tested on the MTs models. If successful, the method could be used clinically to foresee the effectiveness of erlotinib on a patient specific basis, in an ensuing phase 2 SBIR proposal.